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AP Biology
Mrs. Schulte
Experiment 4
January 30-31
Results:
Coffee filter: (because the oil, second from the left, did not produce any effect we left it out of
our report as there was no data to collect)
Water:(furthest on the left)
Paper length 18 cm
Green/yellow line 6 cm
Green line 8 cm
This chart shows the data we collected from putting our solute onto a strip of coffee filter paper
and letting it soak in water. We found the solution (water) traveled all the way up the paper and
the solute (spinach) separated into a green/yellow line and a distinct green line. We measured
the distance the pigments traveled from the original solute line so that we could calculate the Rf
values for both:
Green/yellow Rf value:
6 cm (distance pigment traveled) / 18 cm (distance solution traveled) = .33
Green line 5 cm
Blue line 7 cm
This chart shows the data we collected from putting our solute onto a strip of coffee filter paper
and letting it soak in isopropyl alcohol. We found the solution (the alcohol) traveled all the way
up the paper and the solute (spinach) split into orange, green, yellow, and blue pigments. This
combination gave us the best results out of all of our trials. We measured the distances of each
line to be able to calculate the Rf values:
Yellow line 3 cm
Green line 5 cm
This chart shows the data we collected from placing the solute on a regular filter paper and
letting it soak in isopropyl alcohol. We only put this paper into the alcohol because it is what
gave us the best results prior and we wanted to see if we could get even greater results with
different paper. We found the solution (the alcohol) traveled all the way up the paper and the
solute (spinach) separated into yellow and green pigments. We measured the distance of the
lines of these pigments from the original solute line so we could calculate the Rf values of both.
Green line 10 cm
This chart shows the data we collected from placing the solvent onto a thicker filter paper and
letting it soak in isopropyl alcohol again. We used the alcohol for the same reasons as before;
we found that is gave us the best results prior and wanted to test it on different types of paper.
We found that the solution (the alcohol) traveled all the way up the paper and the solute
(spinach) only separated into a green pigment. We measured its distance from the original
solvnet line so we could find the Rf value:
Green Rf value:
10 cm (distance pigment traveled) / 14.5 cm (distance solution traveled) = .69
Conclusion:
Our hypothesis for this experiment was that we would see the spinach separate into
multiple, distinct colors and that the isopropyl alcohol would cause the greatest results. We
found that our hypothesis was true and the alcohol formed the best results while the oil did
nothing and the water formed little results. Because of this we decided to test the alcohol and
the spinach on different papers thinking thicker filter paper would be the best for this and we
evidently found this not to be true as the thinnest coffee filter paper showed us the greatest
results.
We were able to fulfill our purpose and were able to identify the different compound in
the spinach. On the filter paper with the alcohol, from the bottom up the compounds in the
spinach that separated are chlorophyll b, chlorophyll a, xanthophyll, and carotene. We
successfully used chromatography to separate the pigments in the green plant and were able to
see green, yellow, orange, and a blue line. After performing chromatography we collected
measurements that enabled us to calculate the Rf value for each of the individual pigments in
the plant. With our results we were then able to better understand the relationship and use of
pigments in photosynthesis.
Some sources of error in this lab would include the fact that we did not use actual
chromatography paper instead we used coffee filter paper and then decided to test regular filter
papers of different thicknesses. The chromatography paper would have given us the greatest
results because it is made specifically for this kind of experiment; however, we made due with
what we had. This spinach we used was not fresh spinach as it came from a package and was
most likely packed with preservative chemicals. It was also grown in the winter making it less
“colorful”. If we had used fresh spinach grown in the summer we would have seen greater
results. Another possible source of error could be the fact that we touched the filter paper that
we used for the chromatography and the oils on our fingers could have skewed the data in one
direction or another.
I think it would be interesting to do this lab again but with the proper materials so that we
could get the best possible results and get a greater range of colors/compounds. We could also
test this experiment with different kinds of plants: other green plants and plants of other colors.
For example, would a flower produce the same results?
Works cited:
1) “Separation of Plant Pigments Using Chromatography.” Separation of Plant
Pigments (Chromatography),
www.biologycorner.com/worksheets/plant_pigments.html.